Methods of priming the immunogenic activity of vaccines useful for eliciting a protective immune response

ABSTRACT

Disclosed are methods for vaccine priming, using co-treatment, at a temporally similar or at a previous time, with a priming antibody capable of priming, or enhancing, or potentiating the effects of a vaccine, or vaccine composition. Also disclosed are methods of using this process to prevent or treat disease.

This application is a continuation of U.S. Ser. No. 11/198,323, filedAug. 8, 2005, now U.S. Pat. No. 7,309,490, which is a continuation ofU.S. Ser. No. 10/409,822, filed 9 Apr. 2003, now U.S. Pat. No.6,962,701, which is a continuation of U.S. Ser. No. 09/797,760, filed 2Mar. 2001, now U.S. Pat. No. 6,565,849, which claims the benefit under35 U.S.C. § 119(e) of U.S. Provisional Application Ser. No. 60/186,566,filed 2 Mar. 2000, the disclosure of which is hereby incorporated byreference in their entirety.

FIELD OF THE INVENTION

The present invention relates to a method of priming, enhancing and/orpotentiating the effects of a vaccine using pretreatment with anantibody, including antibodies specific for the antigenic determinantsfrom which the vaccine is derived.

BACKGROUND OF THE INVENTION

The use of vaccines and vaccine compositions is presently a major methodof dealing with the prevention and treatment of disease. However,because such vaccines and vaccine compositions incorporate wholeproteins or polynucleotides or active fragments thereof, theeffectiveness of such treatments can be highly variable. In some cases,highly purified proteins and polypeptides, and fragments thereof,incorporating antigenic determinants characteristic of a microorganism,such as a virus or bacterium, may fail to show sufficiently activeprotection against infection by said organism, possibly because ofstructural or steric shortcomings in the particular polypeptide orfragments used as the basis for the vaccine. Consequently, attempts havebeen made to enhance the effectiveness of such vaccines by addition ofpotentiating agents to the treatment process, especially in cases suchas the treatment of a cancer where said cancer may show unexpectedlyhigh sensitivity to the potentiating agent.

Heretofore, many different procedures have been developed to enhance theactivity of vaccines and vaccine compositions by increasing theimmunogenicity of such compositions. Part of the problem with suchmeasures has been the hit or miss character of these procedures, many ofwhich are based on just trial and error and are not otherwise carefullycalculated to elicit the type of response desired and which makes thevaccine worthwhile. Among the more effective procedures has been the useof adjuvants (such as Freund's complete adjuvant, although the latter isnot useful in treating human patients) as well as the addition of simpleorganic molecules to the vaccine composition.

Many attempts at enhancing the immunogenicity of vaccines have revolvedaround extensive searches for small organic molecules that areeffective, relatively non-toxic and economically advantageous tosynthesize and utilize. Such efforts have met with only spotty success.Such compounds must, of course, be able to mimic the response with morecomplex immunogens, such as lipopolysaccharide of Escherichia coli andwhole polypeptides used in immunogenic compositions. Some agents haveshown promise, for example, the anti-tumor agent of U.S. Pat. No.5,958,980.

Other approaches have involved forming particulate matter, such as byheat-induced aggregation of soluble antigens or by usingself-aggregating antigenic particles (for example, the soluble antigenof hepatitis B virus possesses self-aggregation properties). Suchparticles have been found more useful in attracting immune cells such asmacrophages and inducing immunity in the organism. Other attempts haveemployed more exotic structures such as liposomes, oily droplets, andalum precipitation.

Other methods of enhancing immunogenic activity of vaccines haveincluded attempts to elicit an immunological reaction against an antigenadministered along with a highly potent vaccine such that the latterprovides a kind of adjuvant effect. For example, if a potent immunogenlike killed Bordetella pertussis is administered along with a purifiedprotein or polypeptide, the result is often a stronger immunogenicreaction to the purified protein or polypeptide, possibly due to thefact that the potent immunogen greatly stimulates the production oflymphokines, such as interleukin 4, that possess inter alia strongpolyclonal activating activities.

Other methods have included such processes as slowing down the releaseof the immunogen so as to avoid activating suppressor pathways oravoiding direct intravenous administration, including subcutaneousdevices to slowly release antigen.

Some methods have resorted to genetically engineering the organismsagainst which the immunogenic response is to be directed. For example,the vaccinia virus genome has been engineered to incorporate genes forvarious antigens of different pathogens. [See: Panicali and Paoletti,Proc. Natl. Acad. Sci USA, 79:4927 (1982); Smith et al, Nature, 302:490(1983); Langford et al, Mol. Cell Biol., 6:3191 (1986); also see U.S.Pat. No. 5,879,685 for a highly descriptive survey of pastmethodologies] Other approaches have employed the use of surface activeagents (such as saponin).

However, all of these methods have proven less than completelysatisfactory because each has one or more disadvantages, mostly havingto do with the eliciting of unwanted side reactions involving thestimulation of biological pathways within the recipient organism thatcan have a negative effect on the overall efficacy of the method beingused. For example, alum precipitation often shows unwanted inflammatoryeffects. In addition, co-administration of antigen along with a potentimmunogen is often useful only for a limited number of small peptides.Further, genetically engineering potentially dangerous microorganismsnecessarily engenders safety concerns.

The present invention avoids these disadvantages by providing a means ofpotentiating, or enhancing, or priming the immunogenic effects of avaccine, or vaccine composition, by utilizing a pretreatment stepinvolving a highly specific and high affinity neutralizing antibody toprime the immunogenic reaction and thereby potentiate the effects ofsubsequent administration of the appropriate vaccine, or vaccinecomposition.

BRIEF SUMMARY OF THE INVENTION

The present invention relates to a process for potentiating theimmunogenic reaction of a given vaccine through pretreatment of anorganism, such as a human patient, with a therapeutically effectiveamount of an antibody to prime response to the vaccine.

It is thus an object of the present invention to provide a method forenhancing the immunogenic activity of a vaccine, or vaccine composition,by first administering to an animal, such as a human patient, atherapeutically effective amount or dose of an antibody specific for anantigenic determinant of a cell or microorganism against which saidanimal is to be immunized, followed by administration to the same animalof an effective amount, or dose, of a vaccine, or vaccine composition,comprising at least one antigenic determinant for which the earlieradministered antibody is specific.

It is another object of the present invention to provide a means ofpriming, or enhancing, or potentiating, the immunogenic activity ofvaccines, and vaccine compositions, useful in immunizing againstmicroorganisms, especially bacteria and viruses.

It is a further object of the present invention to provide a method forpriming, enhancing or potentiating the immunogenic activity of vaccines,and vaccine compositions, useful in treating, and or preventing,diseases, especially where said disease afflicts a human patient, orwhere said human is at risk of contracting said disease. Among thediseases to be prevented or treated are those caused by bacteria andviruses, as well as chronic diseases, such as cancer.

DETAILED SUMMARY OF THE INVENTION

The present invention relates to a process for priming, or enhancing, orpotentiating, all of which are considered synonymous when applied to theprocess of the present invention, an immunological response, whereby ananimal is first treated by administration of a sample of an antibodyfollowed at some finite time later by the administration to the sameanimal of a sample of a vaccine, or vaccine composition, whoseimmunogenic activity is to be primed, enhanced, or potentiated.

In accordance with the disclosure herein, the present invention relatesto a process for priming, enhancing, or potentiating immunogenicactivity such as in response to a vaccine, or vaccine composition,comprising:

(a) administering to an animal in need thereof an effective amount of anantibody specific for one or more of the antigenic determinantscharacteristic of said vaccine, or vaccine composition; and

(b) administering to said animal an effective amount of said vaccine, orvaccine composition.

In accordance with the present invention, this process for priming theimmunogenic activity of the vaccine, or vaccine composition, may involveadministering the antibody at some point in time prior to administrationof the vaccine, or vaccine composition, whose immunogenic activity is tobe primed, enhanced or potentiated. Thus, the antibody may beadministered an hour, or a few hours, prior to the administration of thevaccine, or vaccine composition, or said antibody may be administered upto 24 hours prior to said vaccine, or vaccine composition, or saidantibody may be administered up to 48 hours, or up to 72 hours, or evenup to a week, or even longer, prior to administration of said vaccine,or vaccine composition. The antibody and the vaccine, or vaccinecomposition, whose immunogenic activity is to be primed, enhanced orpotentiated may also be administered simultaneously, either separatelyor as part of the same composition, or may even be administered only afew minutes apart. In addition, the vaccine, or vaccine composition, mayeven be administered just prior to administration of the antibody,possible up to several hours or even a day prior thereto. In applyingthe methods disclosed herein, it should be borne in mind that the objectof said invention is to have the antibody and vaccine, or vaccinecomposition, work in sequence to achieve the desired priming, enhancingor potentiating of the underlying immunogenic effect.

As used herein, the term “priming” in its narrowest sense is synonymouswith the terms “enhancing” and “potentiating” and refers to any form ofincrease in the immunogenic activity of a vaccine, or vaccinecomposition, as a result of administering to an animal, such as a human,a therapeutically effective dose of an antibody wherein said antibody isadministered at any time prior to, simultaneous with, or just afteradministration to the same animal of an effective dosage of a vaccine,or vaccine composition, and wherein the antibody is specific for atleast one antigenic determinant characteristic of the active epitopespresent in said vaccine, or vaccine composition.

The present invention is also directed to a process in which either theantibody or the vaccine, or vaccine composition, represents a less thantherapeutically effective dose of the respective antibody or vaccine, orvaccine composition, but together, as a result of the priming process,result in therapeutically effective action in treating the condition tobe ameliorated, whether this be a disease caused by infection, or due tocancer, or some toxic effect due to the exposure of the patient to oneor more toxins. Also in accordance with the present invention, both theantibody and the vaccine, or vaccine composition, may be administered ata less than therapeutically effective dose but, because of the primingeffect, such combination results in effective therapy. Likewise, theantibody and/or the vaccine, or vaccine composition, according to thepresent invention may be administered in one or more (i.e., in a series)of doses, each of which is not by itself therapeutically effective totreat the subject condition or malady, but which in total, because ofthe priming effect, result in therapeutically effective treatment ofsaid condition or malady. The advantage of such a schedule ofadministration might be, inter alia, potentially toxic effects of theantibody, vaccine, or both when given in doses therapeutically effectivewithout the priming effect. Thus, in accordance with the presentinvention, such priming has the advantage of achieving atherapeutically, or prophylactically, effective dosage of either theantibody, or vaccine, or both, but with reduced, or absent, toxiceffects.

In accordance with the disclosure herein, the term “priming” includesenhancement of a normal immune function, such as an immunogenicreaction, as well as the restoration of a depressed immune reaction,such as where the immunogenic activity of the vaccine, or vaccinecomposition, is either inadequate to be of genuine clinical value orelse is simply reduced in effect, or is considered adequate for clinicaluse but is susceptible to enhancement, or increase, in activity.Consequently, the antibody, or antibodies, used according to the presentinvention are essentially co-stimulatory agents, or immunopotentiatingagents, with respect to the vaccine, or vaccine composition, employed inthe claims hereof.

Also in accordance with the present invention, there are providedvaccines, and vaccine compositions, for use herein. Such vaccinescomprise immunogens that themselves may include proteins, nucleic acids,and active fragments thereof, the only real requirement being theability of such substances to elicit an immunogenic response whenintroduced into an animal, especially a human.

The term “active fragment” means a fragment that generates an immuneresponse (i.e., has immunogenic activity) when administered, alone oroptionally with a suitable adjuvant, to an animal, such as a mammal, forexample, a rabbit or a mouse, and also including a human.

As used herein, the terms “portion,” “segment,” and “fragment,” whenused in relation to polypeptides, refer to a continuous sequence ofresidues, such as amino acid residues, which sequence forms a subset ofa larger sequence. For example, if a polypeptide were subjected totreatment with any of the common endopeptidases, such as trypsin orchymotrypsin, the oligopeptides resulting from such treatment wouldrepresent portions, segments or fragments of the starting polypeptide.When used in relation to a polynucleotides, such terms refer to theproducts produced by treatment of said polynucleotides with any of thecommon endonucleases.

In accordance with a further aspect of the invention, a vaccine of thetype hereinabove described is administered for the purpose of preventingor treating a disease, such as an infection, for example, that caused bya microorganism, such as a bacterium or a virus.

A vaccine in accordance with the present invention may include one ormore immunogenic polypeptides or active fragments thereof. Whenemploying more than one polypeptide or active fragment, such as two ormore polypeptides and/or active fragments, these may be used as aphysical mixture or as a fusion of two or more polypeptides or activefragments to form a larger structure or complex, said larger structureor complex having said component polypeptides or fragments linkedtogether either covalently or by some other chemical means, includingelectrostatic attraction, and, where covalently linked, said linkage canbe in the form of a conventional peptide bond, or may involve some othertype of linkage, such as some other type of covalent bond or hydrophobicinteraction, possibly involving linkages utilizing non-peptidesequences, such as linkages that are oligomeric or polymeric in natureand not involving subunits composed of ordinary amino acids. Such afusion fragment or fusion polypeptide may be produced, for example, byrecombinant techniques or by the use of appropriate linkers for fusingpreviously prepared polypeptides or active fragments.

In many cases, the variation in the polypeptide or active fragment is aconservative amino acid substitution, although other substitutions arewithin the scope of the invention.

In accordance with the present invention, a polypeptide variant includesvariants in which one or more amino acids are substituted and/or deletedand/or inserted.

The vaccines finding use in the methods of the present invention alsoinclude polynucleotides, either DNA or RNA, either single, double ortriple stranded, including immunogenically active fragments, segmentsand portions thereof.

In another aspect, the invention relates to passive immunity vaccinesformulated from antibodies against a polypeptide or active fragment of apolypeptide of the present invention. Such passive immunity vaccines canbe utilized, for example, to prevent and/or treat pneumococcalinfections in patients. In this manner, according to a further aspect ofthe invention, a vaccine can be produced from a synthetic or recombinantpolypeptide or an antibody against such polypeptide. Thus, the vaccine,or vaccine composition, may be composed of, or may contain in additionto other antigenic structures, one or more antibodies that are thenprimed by the introduction of an additional antibody, such as where thelatter is introduced prior to the vaccine, or vaccine composition.

Still another aspect the present invention relates to a method of usingone or more antibodies (monoclonal or polyclonal, natural orrecombinant, and regardless of how prepared, i.e., by purification froma natural source, or generated by cloning or by direct chemicalsynthesis), preferably, but not necessarily, specific for one or moreantigenic determinants present in the vaccine, or vaccine compositionselected for use in the methods of the present invention.

Generally, vaccines are prepared as injectables, in the form of aqueoussolutions or suspensions. Vaccines in an oil base are also well knownsuch as for inhaling. Solid forms which are dissolved or suspended priorto use may also be formulated. Pharmaceutically acceptable carriers,diluents and excipients are generally added that are compatible with theactive ingredients and acceptable for pharmaceutical use.

The pharmaceutical compositions useful herein also contain apharmaceutically acceptable carrier, including any suitable diluent orexcipient, which includes any pharmaceutical agent that does not itselfinduce the production of antibodies harmful to the individual receivingthe composition, and which may be administered without undue toxicity.Pharmaceutically acceptable carriers include, but are not limited to,liquids such as water, saline, glycerol and ethanol, and the like,including carriers useful in forming sprays for nasal and otherrespiratory tract delivery or for delivery to the ophthalmic system. Athorough discussion of pharmaceutically acceptable carriers, diluents,and other excipients is presented in REMINGTON'S PHARMACEUTICAL SCIENCES(Mack Pub. Co., N.J. current edition).

Vaccine compositions may further incorporate additional substances tostabilize pH, or to function as adjuvants, wetting agents, oremulsifying agents, which can serve to improve the effectiveness of thevaccine.

Vaccines are generally formulated for parenteral administration and areinjected either subcutaneously or intramuscularly. Such vaccines canalso be formulated as suppositories or for oral administration, usingmethods known in the art, or for administration through nasal orrespiratory routes.

The amount of vaccine sufficient to confer immunity to pathogenicbacteria, viruses, or other microbes is determined by methods well knownto those skilled in the art. This quantity will be determined based uponthe characteristics of the vaccine recipient and the level of immunityrequired. Typically, the amount of vaccine to be administered will bedetermined based upon the judgment of a skilled physician. Wherevaccines are administered by subcutaneous or intramuscular injection, arange of 0.5 to 500 μg purified protein may be given. As useful in thepresent invention, such dosages are commonly sufficient to provide about1 μg, possibly 10 μg, even 50 μg, and as much as 100 μg, up to 500 μg ofimmunogenic protein, or immunogenic polypeptide, or immunogenicallyactive fragments thereof. In addition, more than one such activematerial may be present in the vaccine. Thus, more than one antigenicstructure may be used in formulating the vaccine, or vaccine compositionto use in the methods disclosed herein. This may include two or moreindividually immunogenic proteins or polypeptides, proteins orpolypeptides showing immunogenic activity only when in combination,either quantitatively equal in their respective concentrations orformulated to be present in some ratio, either definite or indefinite.Thus, a vaccine composition for use in the processes disclosed hereinmay include one or more immunogenic proteins, one or more immunogenicpolypeptides, and/or one or more immunogenically active immunogenscomprising antigenic fragments of said immunogenic proteins andpolypeptides, the latter fragments being present in any proportionsselected by the use of the present invention. The exact components, andtheir respective quantities, making up the vaccines, and vaccinecompositions, useful in the methods of the present invention aredetermined, inter alia, by the nature of the disease to be treated orprevented, the severity of such condition where it already exists, theage, sex, and general health of the recipient, as well the personal andprofessional experience and inclinations of the researcher and/orclinician utilizing these methods.

The vaccines, or vaccine compositions, useful in the methods disclosedherein are thus enhanced in their immunogenic activity by co-treatmentwith one or more antibodies, related or unrelated to the antigenicdeterminants present in the vaccine composition. Such enhanced activity,or potentiation, or priming may take any conceivable immunological formof reaction. Thus, such enhancement may comprise any increased, andbeneficial, activity of the immune system resulting from co-treatment ofthe subject with both vaccine, or vaccine composition, and with anantibody as disclosed herein. Such enhanced immunological activity thuscomprises an enhancement of any aspect of immune function, including,but not limited to, any of the protective functions of the immune systemof an animal, such as resistance to infection by a pathogen, includingbacteria, viruses, fungi, rickettsia, mycoplasma, and parasites,regardless of the type of organisms or the etiology of the underlyingdisease. Other protective functions of the immune system that may beaffected by the methods disclosed herein, and thus may realize anenhancement, are the other immune functions that are stimulated by useof vaccines, or vaccine compositions. Thus, for example, vaccines thatmay become available for use against various forms of cancers maylikewise be enhanced by priming with antibodies as disclosed herein.Thus specific resistance can be augmented by use of immunopotentiatorsin conjunction with some form of antigen as in a vaccine employing, forexample, attenuated viruses, or even cancer cells. Further, suchapplications can be used not only to induce resistance but also toinduce tolerance, such as where antigens are used in allergy or possiblyeven an auto-immune disease.

Use of the methods of the present invention may thus be eithertherapeutic or prophylactic in nature. Further, the methods disclosedaccording to the present invention may have unique value to specialgroups of individuals, such as the elderly and children, where use oflarge amounts of vaccines may be dangerous or ill advised. Here, throughpriming with antibody, smaller amounts of vaccines may be required toproduce an effect in the same range as that seen with much larger doses,thereby avoiding unwanted side effects or overreaction to theimmunogenic nature of the vaccines being administered. Of course, insuch groups as the elderly, conditions such as infection, auto-immunity,and cancer are more common.

As already described, the timing, dosage and route of administration ofthe vaccines, or vaccine compositions, as well as the antibodies usefulin the present invention are subject to wide variation yet may becritical in determining whether the resulting response thereto is to bepositive or negative. Thus, the methods disclosed herein may be capableof inhibiting immune responses as easily as enhancing them given theright set of conditions. Consequently, the priming procedures describedherein may, with judicious alteration, be used for immunosuppressiveaction as well as enhancement. The former may realize some value whenapplied against allergy, auto-immunity and transplantation.

The immunoenhancement, or priming, resulting from the methods disclosedherein may also affect other aspects of the immune system. Thus, theymay produce an enhancement of either the humoral or the cellularresponse, they may stimulate the action of T lymphocytes, since thelatter have surface receptors keenly sensitive to antigenic fragmentsmaking up some of the vaccines, or vaccine compositions, useful in thepresent invention, with the priming being triggered by priorintroduction of antibodies as disclosed herein. Thus, Immune responsesorchestrated by T-lymphocytes provide a model for the priming effectforming the basis of the present invention in that the actions of such Tlymphocytes, together with antigen presenting cells and majorhistocompatibility complex (MHC), show the value of co-stimulation as ameans of realizing immune function.

Regardless of the mechanism of action of the methods disclosed herein,it is clear that the present invention provides a unique means for thetreatment of diseases where there is a defect in the immune systemand/or an ineffective host defense mechanism, or to enhance activity ofthe immune system above normal levels.

Thus, the methods disclosed herein also find use in the treatment ofanimals showing various types of immunodeficiency, wherein said animal,such as a human patient, has a deficient or defective immune system.This condition can takes the form of inadequate production of antibodiesby the B lymphocytes when presented with foreign antigens as well as adecreased ability to produced T lymphocytes for mounting of a cellulardefense. Thus, the methods disclosed herein have the effect ofameliorating such conditions by priming the immune response elicited bythe vaccines, or vaccine compositions, use herein when coupled with theeffects or prior administration of one or more antibodies, oftenspecific for the antigenic target of the vaccination procedure, such asan invading pathogen. Thus, the methods of the present invention canpartly make up for the presence of immunodeficiency by providing asource of antibody for treatment or prevention of a particular diseasecondition.

In applying the methods of the present invention, it is necessary toprovide an effective amount of the vaccines, or vaccine compositions,used therein. By an “effective amount” is meant the amount of thevaccine, or vaccine composition, that will produce a protectiveimmunogenic effect when coupled with prior administration, orco-administration, of an antibody. It should be noted that the vaccinecompositions useful in the methods of the present invention maythemselves contain an antibody but that said antibody, if present, isconsidered separate from the “priming” antibody essential to the methodsdisclosed herein. Thus, the antibodies used in the present inventionhave the effect of priming the vaccine, or vaccine composition,regardless of the make-up of said vaccine, or vaccine composition.

A further important component of the methods of the present invention isthe use of a “priming” step, comprising administration of one or moreantibodies to an animal, especially a human patient, prior to, alongwith, or just after the administration of a vaccine, or vaccinecomposition. Because the immune response normally elicited by theadministration of a vaccine, or vaccine composition, will commonlyrequire a period of time, or lag period, before the immune response hasreached effective levels, the antibody priming step may be employed evenup until just after administration of the vaccine, or vaccinecomposition. Such administration can thus occur at any time up untiljust after administration of the vaccine, or vaccine composition, andcertainly at the time periods already provided above, even as much as aweek prior to vaccine administration. Furthermore, the administration ofthe priming antibody, or antibodies, within the methods of the presentinvention need in no way be limited to administration of a singleantibody, nor be limited to just one administration of one or more suchpriming antibodies. Thus, the priming antibodies, as used herein, mayinclude any form of antibody, including, but not limited to, monoclonal,polyclonal, recombinant, wholly synthetic, grafted, chimeric,bispecific, single chain, heavy chain, light chain, dimeric, tetrameric,and any active fragments of any of these, such as Fab fragments.

In one embodiment of the present invention, the antibody, or antibodies,used for the priming step (step (a) in the procedure described above)will exhibit specificity toward one or more, but at least one, antigenicdeterminant present in the vaccines, or vaccine compositions, useful inpracticing the invention. However, depending on the mechanism ofpriming, such as by the stimulation of lymphokines or other cytokines,or other wholly different chemical agents, or by whatever means, thepresent invention can readily be practiced using antibodies whosespecificity is wholly different from that of the antigenic determinantspresent in the vaccine, or vaccine composition, whose immunogenicactivity is to be increased.

In addition, the antibodies useful in practicing the present inventionneed not necessarily have any effect on the immunogenic activity of thevaccines, or vaccine compositions, useful for the present invention.Instead, the antibodies useful in practicing the invention, and thevaccines, or vaccine compositions, useful in practicing the inventionmay, when given together, or in some other temporal sequence, have asynergistic effect, such that the sum of the immunological effects ofthe antibody, or antibodies, and the vaccine, or vaccine composition,when administered separately is less that that realized when the primingprocess of the present invention is employed.

In accordance with the present invention, the success of the primingeffect will depend on the dosage and timing of the administration of theantibody, or antibodies, used to achieve the priming effect. As usedherein, the antibody, or antibodies, are commonly administeredintravenously, or intramuscularly, or by nasal inhalation, as well as byother convenient routes and are commonly administered in dosages rangingfrom 5 to 20 mg/kg of body weight of the recipient animal, or from 10 to20 mg/kg, in some cases about 15 mg/kg of body weight.

A further aspect of the present invention provides for the use of thepriming procedure disclosed herein for the treatment and/or prophylaxisof acute and chronic microbial infections.

In accordance with the present invention, such microbial infectionscommonly include viral infections and bacterial infections. Examples ofacute viral infections against which the priming therapy, andprophylaxis, of the present invention finds use includes: respiratorysyncytial virus (RSV), herpes viruses, influenza viruses, parainfluenzaviruses (PIV), adenoviruses, coxsakie viruses, picorna viruses,rotaviruses, heptatis A virus (HAV), hepatitis B virus (HBV), hepatitisC virus (HCV), hepatitis D virus (HDV), hepatitis E virus (HEV),influenza A virus, and influenza B virus, rubellavirus, measles virus,pox viruses, papilloma viruses, and enteroviruses, arenavirus,rhinoviruses, poliovirus, Newcastle disease virus, rabies virus,arboviruses. The methods of the present invention also find use againstrecombinant forms of these viruses where such are to be studied in aresearch setting. For example, the antibodies disclosed in U.S. Pat. No.6,824,307 are particularly useful for treating respiratory syncytialvirus (as well as the asthma and extended periods of wheezing that oftenpersist after infection by this virus).

Of the DNA viruses, those of the herpes group are responsible for manyof the viral infections in humans. This viral group includes: herpessimplex virus (HSV), types 1 and 2, varicella zoster virus (VZV),cytomegalovirus (CMV); Epstein-Barr virus (EBV) and human herpes virus 6(HHV6). HSV 1 and HSV 2 are some of the most common infectious agents ofman.

In a specific embodiment of the present invention the priming antibodyis an anti-RSV antibody specific for the F antigen of RSV and thevaccine is an anti-RSV vaccine comprising one or more epitopes of the Fantigen of RSV (for example, the attenuated RSV vaccine of U.S. Pat. No.5,993,824 or vaccines containing mutant forms of this organism such asthat disclosed in U.S. Pat. No. 5,932,222).

In cases where the methods of the invention are to be employed againstbacterial infections, or the prevention thereof, the bacteria willcommonly include E. coli, pneumococcus pneumonii andpneumocystiscarinii.

The antibodies, as well as vaccines, useful in practicing the presentinvention may be administered to a human recipient by a route selectedfrom oral, parenteral (including subcutaneous, intradermal,intramuscular and intravenous), rectal and inhalation. The size of aneffective dose of a compound will depend upon a number of factorsincluding the identity of the recipient, the type of immunopotentiationinvolved, the severity of the condition to be treated and the route ofadministration, and will ultimately be at the discretion of theattendant physician.

For each of the aforementioned means of administration of antibodies tosaid patient for the purposes of priming subsequent vaccineadministrations, an effective dose of said antibody, or antibodies, willgenerally be in the range 0.5 to 30 mg/kg bodyweight of the humanrecipient, on a per daily regimen, preferably in the range 5 to 20 mg/kgbodyweight per day and most preferably in the range 10 to 15 mg/kgbodyweight per day; an optimum dose is 15 mg/kg bodyweight per day,these doses being for human usage only.

The particular dose to be administered may be given as a series ofdoses, for example, as several sub-doses given over the course of a day,such as between two and four sub-doses administered at appropriateintervals throughout the time course of administration, or may beadministered as a single dose, with the number of doses, the dosage ofeach dose, and the time course depending intimately on the nature of thevaccine, or vaccine composition, the condition of the patient, and thenature of the disease to be either treated or prevented. Thus wherethree sub-doses are employed each will generally lie in the range 0.1 mgto 10 mg/kg of body weight, preferably 0.5 mg to 5 mg/kg body weight,and most preferably 1 mg to 5 mg/kg bodyweight with an optimum of 5mg/kg bodyweight. A daily dose for a human weighing of the order of 50kg will thus generally lie in the range of about 450 mg. Optimally ahuman daily dose is 450 mg.

In one embodiment, wherein the disease condition to be treated orprevented is respiratory syncytial virus, the treatment may call for arange of about 450 mg as a daily dose, given once a month for a periodof about 5 months. The anti-RSV vaccine to be primed in such a procedurewould then be administered either at the end of the 5 month period, orcould be administered in monthly intervals starting at the time of thesecond dose of anti-RSV antibody, or some other regimen could be used.Alternatively, administration of the vaccine, or vaccine compositioncould be carried out simultaneously with each of the antibodyadministrations, or possibly each of the administrations of vaccine, orvaccine composition, could be given on the day following theadministration of antibody, or any other regimen devised by the sounddiscretion of the clinician after review of the needs of the particularpatient as well as considerations of the tolerance of said patient tovarying dosages of the vaccine, or vaccine composition, as well as theantibody, or antibodies, to be used.

For an anti-RSV antibody, the desired dose is preferably presented as asingle dose, at monthly intervals, over a 5 month period.

It should be reiterated that the process for vaccine priming, asdisclosed herein, is a general procedure that can operate to the benefitof any vaccine, or vaccine composition, and employing any antibody.Thus, the antibody use to prime a given vaccine, or vaccine composition,can be selected independently of the particular vaccine, or vaccinecomposition, and does not have to be an antibody specific for anantigenic determinant of the vaccine. although in such cases the primingeffect may work more effectively.

The formulations of the antibodies useful for the processes describedherein, include those suitable for parenteral (including subcutaneous,intradermal, intramuscular and intravenous) and nasal or pulmonaryadministration although the most suitable route may depend upon the ageand condition of the recipient. Such formulations may be presented inunit dosage form and may be prepared by any methods well known in thepharmaceutical arts. Of course, all such methods include the conditionthat the antibodies used, however formulated, maintain their specificityfor the antigenic determinants that served as the basis for selection.Thus, because of the polypeptide nature of the antibodies useful in themethods of the invention, any formulation that fails to maintain theoverall structural integrity of the antibody protein molecules willlikely prove unsuccessful in carrying out the processes according to thepresent invention. However, formulations known to maintain suchstructural integrity are well known in the art and include such carriersas saline and glycerol. In general the formulations are prepared byuniformly and intimately bringing into association the active ingredientwith liquid carriers or finely divided solid carriers or both.

Formulations for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain antioxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which may include suspending agents and thickeningagents. The formulations may be presented in unit-dose or multi-dosecontainers, for example sealed ampoules and vials, and may be stored ina freeze-dried (lyophilized) condition requiring only the addition ofthe sterile liquid carrier, for example water for injections,immediately prior to use. Solutions and suspensions suitable forinjection may be prepared just prior to use.

Formulations suitable for pulmonary administration via the buccal cavityare presented such that particles containing the active ingredient anddesirably having a diameter of under 10 μm are delivered into thebronchial tree of the recipient.

As one possibility such formulations are in the form of fine powdersthat may conveniently be presented either in an easily pierced capsule,such as a gelatin, for use in an inhalation device, or alternatively asa formulation comprising active ingredient, a suitable liquid propellantand optionally other ingredients such as surfactant and/or a soliddiluent. So-called “self-propelling” formulations of this nature mayalso be employed wherein the active ingredient is dispensed in the formof droplets of a solution or suspension.

Such formulations are analogous to those well known in the art and maybe prepared by established procedures. Such formulations areadvantageously presented in a container provided with either amanually-operable or automatically functioning valve having the desiredspray characteristics and having some type of valve structure fordelivering a metered amount or fixed volume, for example 50 to 100microliters, upon each operation thereof.

Solutions of the antibodies useful for the methods disclosed herein mayalso be maintained in the form of a solution for use in an atomiser ornebuliser whereby an accelerated airstream, whether by ultrasonicagitation or some other means, is employed to produce a fine dropletmist for inhalation by the patient. Such methods can also be applied tonasal administration or, for that matter, to any type of introduction ofsuch antibodies via the respiratory system.

More specifically, formulations for nasal administration should have aparticle diameter in the range 10 to 200 microns to enable retention inthe nasal cavity, often achieved through use of a powder of the properparticle diameters. Of course, dosage levels must be adjustedaccordingly.

Preferred unit dosage formulations are those containing an effectivedose, as hereinabove recited, or an appropriate fraction thereof, of theactive antibody.

The present invention is also directed to a vaccine in which apolypeptide or active fragment thereof is delivered or administered inthe form of a polynucleotide encoding the polypeptide or activefragment, whereby the polypeptide or active fragment is produced invivo. The polynucleotide may be included in a suitable expression vectorand combined with a pharmaceutically acceptable carrier.

In addition, polypeptides can be used as immunogens to stimulate theproduction of antibodies for use in passive immunotherapy, for use asdiagnostic reagents, and for use as reagents in other processes such asaffinity chromatography.

The present invention also relates to methods of treating or preventinga disease by carrying out the steps described above on an animal at riskof said disease, or afflicted with said disease. Such animal is commonlya human being. Such disease can include that caused by a pathogen,including diseases caused by any and all of the pathogens alreadyenumerated, as well as chronic diseases, such as cancer.

In carrying out the procedures of the present invention it is of courseto be understood that reference to particular buffers, media, reagents,cells, culture conditions and the like are not intended to be limiting,but are to be read so as to include all related materials that one ofordinary skill in the art would recognize as being of interest or valuein the particular context in which that discussion is presented. Forexample, it is often possible to substitute one buffer system or culturemedium for another and still achieve similar, if not identical, results.Those of skill in the art will have sufficient knowledge of such systemsand methodologies so as to be able, without undue experimentation, tomake such substitutions as will optimally serve their purposes in usingthe methods and procedures disclosed herein.

In practicing the methods disclosed according to the present invention,it should be borne clearly in mind that other and different embodimentsof these processes, in addition to those specifically disclosed herein,will no doubt suggest themselves to those of skill in the relevant art.

1. A process for priming the immunogenic activity of a vaccine,comprising: (a) administering to an animal in need thereof an effectiveamount of an antibody specific for one or more of the antigenicdeterminants characteristic of said vaccine useful against a toxin andwhose immunogenic activity is to be primed; and (b) administering tosaid animal an effective amount of the vaccine, of step (a); whereinsaid vaccine comprises at least one antigenic determinant for which theantibody of step (a) is not specific.
 2. The process of claim 1, whereinstep (a) is carried out at least 24 hours before step (b).
 3. Theprocess of claim 1, wherein step (a) is carried out at least 1 weekbefore step (b).
 4. The process of claim 1, wherein the antibody isspecific for more than one antigenic determinant present in the vaccine.5. The process of claim 1, wherein the effective amount of antibody isin the range of 5 to 20 mg/kg of body weight.
 6. The process of claim 1,wherein the effective amount of vaccine is between about 5 μg per kg ofbody weight to about 1 gram per kg body weight.
 7. A process for primingthe immunogenic activity of a vaccine, comprising: (a) administering toan animal in need thereof an effective amount of an antibody specificfor one or more of the antigenic determinants characteristic of saidvaccine useful against a bacterium and whose immunogenic activity is tobe primed; and (b) administering to said animal an effective amount ofthe vaccine, of step (a); wherein said vaccine comprises at least oneantigenic determinant for which the antibody of step (a) is notspecific.
 8. The process of claim 7, wherein step (a) is carried out atleast 24 hours before step (b).
 9. The process of claim 7, wherein step(a) is carried out at least 1 week before step (b).
 10. The process ofclaim 7, wherein the antibody is specific for more than one antigenicdeterminant present in the vaccine.
 11. The process of claim 7, whereinthe effective amount of antibody is in the range of 5 to 20 mg/kg ofbody weight.
 12. The process of claim 7, wherein the effective amount ofvaccine is between about 5 μg per kg of body weight to about 1 gram perkg body weight.
 13. A process for priming the immunogenic activity of avaccine, comprising: (a) administering to an animal in need thereof aneffective amount of an antibody specific for one or more of theantigenic determinants characteristic of said vaccine which elicits animmunogenic response and whose immunogenic activity is to be primed; and(b) administering to said animal an effective amount of the vaccine, ofstep (a); wherein said vaccine comprises at least one antigenicdeterminant for which the antibody of step (a) is not specific.
 14. Theprocess of claim 13, wherein step (a) is carried out at least 24 hoursbefore step (b).
 15. The process of claim 13, wherein step (a) iscarried out at least 1 week before step (b).
 16. The process of claim13, wherein the antibody is specific for more than one antigenicdeterminant in the vaccine.
 17. The process of claim 13, wherein theeffective amount of antibody is in the range of 5 to 20 mg/kg of bodyweight.
 18. The process of claim 13, wherein the effective amount ofvaccine is between about 5 μg per kg of body weight to about 1 gram perkg body weight.
 19. The process of claim 13, wherein the vaccine elicitsan immunogenic response to a cancer antigen.
 20. The process of claim 7,wherein the bacterium is selected from the group consisting of E. coli,pneumococcus, and pneumocystis.
 21. The process of claim 1, wherein theanimal is a human being.
 22. The process of claim 7, wherein the animalis a human being.
 23. The process of claim 13, wherein the animal is ahuman being.
 24. The process of claim 1, wherein the effective amount ofantibody is in the range of 10 to 20 mg/kg of body weight.
 25. Theprocess of claim 7, wherein the effective amount of antibody is in therange of 10 to 20 mg/kg of body weight.
 26. The process of claim 13,wherein the effective amount of antibody is in the range of 10 to 20mg/kg of body weight.
 27. The process of claim 1, wherein step (a) iscarried out at least 48 hours before step (b).
 28. The process of claim1, wherein step (a) is carried out at least 72 hours before step (b).29. The process of claim 7, wherein step (a) is carried out at least 48hours before step (b).
 30. The process of claim 7, wherein step (a) iscarried out at least 72 hours before step (b).
 31. The process of claim13, wherein step (a) is carried out at least 48 hours before step (b).32. The process of claim 13, wherein step (a) is carried Out at least 72hours before step (b).
 33. The process of claim 1, wherein the effectiveamount of vaccine is between about 100 μg per kg of body weight to about1 gram per kg body weight.
 34. The process of claim 1, wherein theeffective amount of vaccine is between about 1 per kg of body weight toabout 1 gram per kg body weight.
 35. The process of claim 1, wherein theeffective amount of vaccine is between about 10 mg per kg of body weightto about 1 gram per kg body weight.
 36. The process of claim 7, whereinthe effective amount of vaccine is between about 100 μg per kg of bodyweight to about 1 gram per kg body weight.
 37. The process of claim 7,wherein the effective amount of vaccine is between about 1 mg per kg ofbody weight to about 1 gram per kg body weight.
 38. The process of claim7, wherein the effective amount of vaccine is between about 10 mg per kgof body weight to about 1 gram per kg body weight.
 39. The process ofclaim 13, wherein the effective amount of vaccine is between about 100μg per kg of body weight to about 1 gram per kg body weight.
 40. Theprocess of claim 13, wherein the effective amount of vaccine is betweenabout 1 mg per kg of body weight to about 1 gram per kg body weight. 41.The process of claim 13, wherein the effective amount of vaccine isbetween about 10 mg per kg of body weight to about 1 gram per kg bodyweight.